清热利湿饮通过调控miR-17-5p抑制人角质形成细胞STAT3通路的作用研究

Qingre Lishi Recipe suppresses STAT3 signaling through upregulating miR-17-5p in keratinocytes cells

目的确定清热利湿饮对人角质形成细胞JAK-STAT3信号通路的影响,探讨抑制JAK-STAT3信号通路在清热利湿饮治疗银屑病机制的作用。方法Western blot检测细胞蛋白的表达,实时定量PCR检测JAK mRNA的表达,荧光素酶报告基因检测STAT3的转录以及JAK 3‘UTR的活性。结果清热利湿饮显著降低人角质形成细胞株HaCaT细胞内STAT3磷酸化水平以及STAT3的转录功能;进一步研究显示清热利湿饮可通过转录后水平降低JAK1表达抑制STAT3的磷酸化;实时定量PCR和荧光素酶报告基因分析实验结果显示清热利湿饮能够促进miR-17-5p的表达,进而抑制JAK13‘UTR活性,抑制JAK1的翻译,降低JAK1的表达。结论清热利湿饮能通过促进miR-17-5p的表达,抑制人角质形成细胞内JAK1的表达,进而抑制STAT3的功能。

Objective In order to characterize the effect of Qingre Lishi recipe on STAT3 in HaCaT cells.Methods Cells were treated with Qingre Lishi recipe and protein expression was detected by western blot,mRNA expression was measured by real-time PCR,and STAT3 transcription and JAK13‘UTR activity was measured by luciferase reporter assay.Results Qingre Lishi recipe significantly decreases the levels of phosphorylated STAT3 and STAT3 transcription activity in HaCaT cells.We further showed that Qingre Lishi recipe downregulated phosphorylated STAT3 levels through decreasing JAK1 expression at posttranscriptional level.Mechanically,Qingre Lishi recipe could upregulate miR-17-5 p levels and decrease JAK13‘UTR activity,which in turn inhibits JAK1 translation.Conclusion Qingre Lishi recipe could increase miR-17-5 p levels in HaCaT cells,which targets JAK13‘UTR and decreased JAK1 protein levels,therefore downregulates STAT3 activity.